ELISA Kit for Toll Like Receptor 4 (TLR4)

CD284; TOLL; HToll

Specificity

This assay has high sensitivity and excellent specificity for detection of Toll Like Receptor 4 (TLR4).
No significant cross-reactivity or interference between Toll Like Receptor 4 (TLR4) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Toll Like Receptor 4 (TLR4) and the recovery rates were calculated by comparing the measured value to the expected amount of Toll Like Receptor 4 (TLR4) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 92-99 96
EDTA plasma(n=5) 78-104 88
heparin plasma(n=5) 80-103 94

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Toll Like Receptor 4 (TLR4) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Toll Like Receptor 4 (TLR4) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Toll Like Receptor 4 (TLR4) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 79-101% 94-103% 95-102% 84-99%
EDTA plasma(n=5) 78-89% 87-95% 84-98% 83-90%
heparin plasma(n=5) 87-104% 82-89% 91-98% 78-95%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Chinese Medical Journal Expression of soluble Toll-like receptors in pleural effusions CMJ: 2010823483999506297
Journal of Maternal-Fetal and Neonatal Medicine Amniotic fluid soluble Toll-like receptor 4 in pregnancies complicated by preterm prelabor rupture of the membranes Informa: 14767058.2011.626821
PLoS One Baicalein Reduces Airway Injury in Allergen and IL-13 Induced Airway Inflammation PubMed: PMC3639905
Clin Res Hepatol Gastroenterol. The association between precancerous gastric lesions and serum pepsinogens, serum gastrin, vascular endothelial growth factor, serum interleukin-1 Beta, serum toll-like receptor-4 levels and Helicobacter pylori Cag A status Pubmed: 23137754
Journal of Clinical Microbiology Toll-Like Receptor 2 in Serum: a Potential Diagnostic Marker of Prosthetic Joint Infection? Pubmed: 24478497
Surgical Endoscopy CO2 pneumoperitoneum increases secretory IgA levels in the gut compared with laparotomy in an experimental animal model. Pubmed: 24442682
Experimental and therapeutic medicine Soluble toll-like receptor 4 reversed attenuating effect of Chinese herbal Xiao-Qing-Long-Tang on allergen induced nerve growth factor and thymic stromal lymphopoietin Pubmed: 24223644
Free Radical Biology and Medicine Involvement of nitric oxide with activation of Toll-like receptor 4 signaling in mice with dextran sodium sulfate-induced colitis Pubmed:24992835
Respiratory Virology and Immunogenicity Advances in Experimental Medicine and Biology Cytokines and Toll-Like Receptors in the Immune Response to Influenza Vaccination Pubmed:25248345
Molecular and Cellular Biochemistry Beta-arrestin-2 negatively modulates inflammation response in mouse chondrocytes induced by 4-mer hyaluronan oligosaccharide Pubmed:25318610
Journal of Ginseng Research Effects of Korean Red Ginseng (Panax ginseng), urushiol (Rhus vernicifera Stokes), and probiotics (Lactobacillus rhamnosus R0011 and Lactobacillus acidophilus R0052) on the gut–liver axis of alcoholic liver disease Pubmed:25378990
BMC Immunology The discriminative capacity of soluble Toll-like receptor (sTLR)2 and sTLR4 in inflammatory diseases Pubmed:25406630
Gastroenterology Research and Practice Procoagulatory State in Inflammatory Bowel Diseases Is Promoted by Impaired Intestinal Barrier Function Pubmed:25767508
Eur J Gastroenterol Hepatol Efficacy of rifaximin on circulating endotoxins and cytokines in patients with nonalcoholic fatty liver disease PubMed: 26043290
Pathol Biol (Paris) Cathelicidin related antimicrobial peptide, laminin, Toll-like receptors and chemokines levels in experimental hypersensitivity pneumonitis in mice PubMed: 25834936
Journal of Food Science  Effects of Hyriopsis cumingii Polysaccharides on Mice Immunologic Receptor, Transcription Factor, and Cytokine Pubmed:27061742
Plos One Soluble Toll-Like Receptors 2 and 4 in Cerebrospinal Fluid of Patients with Acute Hydrocephalus followingAneurysmal Subarachnoid Haemorrhage. Pubmed:27223696
Open Forum Infect Dis The Effect of Rosuvastatin on Markers of Immune Activation in Treatment-Naive Human Immunodeficiency Virus-Patients Pubmed:26835476
Animal Feed Science and Technology Impact of Saccharomyces cerevisiae fermentation product and subacute ruminal acidosis on production, inflammation, and fermentation in the rumen and hindCavia (Guinea pig )t of dairy cows science:S0377840115300493
european journal of pharmacology Garlic active constituent s-allyl cysteine protects against lipopolysaccharide-induced cognitive deficits in the rat: Possible involved mechanisms. pubmed:27915041
european journal of pharmacology S-allyl cysteine ameliorates cognitive deficits in streptozotocin-diabetic rats via suppression of oxidative stress, inflammation, and acetylcholinesterase. pubmed:27887948
Psychopharmacology (Berl) Ellagic acid ameliorates learning and memory deficits in a rat model of Alzheimer's disease: an exploration of underlying mechanisms. pubmed:28303372
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biomedicine & pharmacotherapy Acetyl-l-carnitine protects dopaminergic nigrostriatal pathway in 6-hydroxydopamine-induced model of Parkinson's disease in the rat. pubmed:28199883
Journal For Neurovirology Soluble TLR2 and 4 concentrations in cerebrospinal fluid in HIV/SIV-relatedneuropathological conditions. pubmed:27882497
Acta Orthopaedica Toll-like receptors and their soluble forms differ in the knee and thumb basal osteoarthritic joints pubmed:28093922
Environmental Toxicology and Pharmacology Ameliorative potential of linagliptin and/or calcipotriol on bleomycin-induced lung fibrosis: In vivo and in vitro study pubmed:28192751
Inflammation Toll-Like Receptor 4 Inhibition Improves Oxidative Stress and Mitochondrial Health in Isoproterenol-Induced Cardiac Hypertrophy in Rats pubmed:28690610
Comparative Immunology, Microbiology and Infectious Diseases Probiotic Lactobacillus acidophilus bacteria or synthetic TLR2 agonist boost the growth of chicken embryo intestinal organoids in cultures comprising epithelial cells and myofibroblasts DOI: 10.1016/j.cimid.2017.06.002
Comparative Immunology, Microbiology and Infectious Diseases Probiotic Lactobacillus acidophilus bacteria or synthetic TLR2 agonist boost the growth of chicken embryo intestinal organoids in cultures comprising epithelial cells and myofibroblasts 10.1016/j.cimid.2017.06.002
Microbial Pathogenesis Protective effects of sinomenine against LPS-induced inflammation in piglets. pubmed:28757275
Cytokine Soy isoflavone genistein attenuates lipopolysaccharide-induced cognitive impairments in the rat via exerting anti-oxidative and anti-inflammatory effects pubmed:29102164
Biomedicine & pharmacotherapy Berberine ameliorates intrahippocampal kainate-induced status epilepticus and consequent epileptogenic process in the rat: Underlying mechanisms pubmed:28061403
Frontiers in Immunology The soluble Form of Toll-like receptor 2 is elevated in serum of Multiple sclerosis Patients: a novel Potential Disease Biomarker Pubmed:29593720
Experimental Gerontology Specific alterations in the circulating levels of the SIRT1, TLR4, and IL7 proteins in patients with dementia Pubmed:30071285
Neurotoxicity Research Protective Effects of Fibroblast Growth Factor 21 Against Amyloid-Beta-Induced Toxicity in SH-SY5Y Cells Pubmed:29869772
European Journal of Pharmacology Rutin, a quercetin glycoside, alleviates acute endotoxemic kidney injury in C57BL/6 mice via suppression of inflammation and up-regulation of antioxidants and SIRT1 Pubmed:29920283
Environmental Toxicology and Pharmacology Crocin reverses unilateral renal ischemia reperfusion injury-induced augmentation of oxidative stress and toll like receptor-4 activity Pubmed:29625388
Physiology & Behavior  Linagliptin potentiates the effect of l-dopa on the behavioural, biochemical and immunohistochemical changes in experimentally-induced Parkinsonism: Role of toll … Pubmed:29410018
International Journal of Gynecology & Obstetrics Increased serum levels of high mobility group protein B1 and calprotectin in pre‐eclampsia Pubmed:29569400
Journal of Maternal-Fetal & Neonatal Medicine Increased TLR4 and TREM-1 expression on monocytes and neutrophils in preterm birth: further evidence of a proinflammatory state Pubmed:29534643
ТLR4-РІВНІ У ХВОРИХ НА ПОДАГРУ: ЗВ'ЯЗОК З АКТИВНІСТЮ ЗАХВОРЮВАННЯ :
Journal of Medical Virology Combined effect of anti‐high‐mobility group box‐1 monoclonal antibody and peramivir against influenza A virus‐induced pneumonia in mice Pubmed: 30281823
Metabolic Brain Disease Berberine ameliorates lipopolysaccharide-induced learning and memory deficit in the rat: insights into underlying molecular mechanisms Pubmed: 30456649
Experimental and Therapeutic Medicine The HMGB1‑RAGE/TLR‑TNF‑α signaling pathway may contribute to kidney injury induced by hypoxia
Immunopharmacology and immunotoxicology Protective effect of sesamin in lipopolysaccharide-induced mouse model of acute kidney injury via attenuation of oxidative stress, inflammation, and apoptosis Pubmed: 30488751
International Immunopharmacology S-allyl cysteine protects against lipopolysaccharide-induced acute kidney injury in the C57BL/6 mouse strain: Involvement of oxidative stress and inflammation Pubmed: 30665040
Biomedicine & Pharmacotherapy Opuntioside, opuntiol and its metallic nanoparticles attenuate adjuvant-induced arthritis: Novel suppressors of Toll-like receptors-2 and-4
International Journal of Obesity Circulating microbiota-derived metabolites: a “liquid biopsy? Pubmed: 31388096
Scientific Reports Deregulation of TLR4 signaling pathway characterizes Bicuspid Aortic valve syndrome Pubmed: 31363123
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